Human GSTK1 knockout HEK-293T cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human GSTK1 knockout HEK-293T cell line (AB266112)
Lanes 1-4 : Merged signal (red and green). Green - ab52759 observed at 27 kDa. Red - loading control ab8245 observed at 36 kDa.
ab52759 Anti-GSTK1 antibody [EP1938Y] was shown to specifically react with GSTK1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266112 (knockout cell lysate ab257461) was used. Wild-type and GSTK1 knockout samples were subjected to SDS-PAGE. ab52759 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-GSTK1 antibody [EP1938Y] (<a href='/en-us/products/primary-antibodies/gstk1-antibody-ep1938y-ab52759'>ab52759</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
GSTK1 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human GSTK1 knockout HEK-293T cell line (ab266112)
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
Daudi cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 25 kDa
Observed band size: 27 kDa
false
- WB
Lab
Western blot - Human GSTK1 knockout HEK-293T cell line (AB266112)
Lanes 1-4 : Merged signal (red and green). Green - ab134173 observed at 27 kDa. Red - loading control ab8245 observed at 36 kDa.
ab134173 Anti-GSTK1 antibody [EPR1939] was shown to specifically react with GSTK1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266112 (knockout cell lysate ab257461) was used. Wild-type and GSTK1 knockout samples were subjected to SDS-PAGE. ab134173 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-GSTK1 antibody [EPR1939] (<a href='/en-us/products/primary-antibodies/gstk1-antibody-epr1939-ab134173'>ab134173</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
GSTK1 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human GSTK1 knockout HEK-293T cell line (ab266112)
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
Daudi cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 25 kDa
Observed band size: 27 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human GSTK1 knockout HEK-293T cell line (AB266112)
Homozygous : 1 bp insertion in exon 1
Complementary Products
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSTK1 antibody [EP1938Y] (AB52759)](https://content.abcam.com/products/images/gstk1-antibody-ep1938y-ab52759--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img63077.jpg)
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
GSTK1 is part of the cellular defense mechanism against xenobiotic and oxidative stress. This enzyme participates in the neutralization of harmful compounds by facilitating their conjugation with glutathione a powerful antioxidant. GSTK1 functions independently and does not form part of a larger protein complex. Its activity is important in maintaining cellular homeostasis and preventing damage caused by reactive oxygen species.
Pathways
GSTK1 has critical involvement in glutathione metabolism and xenobiotic detoxification pathways. It interacts functionally with proteins like GSTM1 which are also part of the glutathione S-transferase family enhancing the cell's ability to detoxify harmful substances. GSTK1 helps to modulate the cellular response to oxidative stress through these pathways ensuring protection against toxicity.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com